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目的观察核受体Nur77在不同发育阶段小鼠睾丸的表达,并探讨其对睾酮合成的作用。方法用1周龄的ICR雄性小鼠作为幼年期组(相当于人类幼年期,n=6),4周龄的ICR雄性小鼠作为性发育前组(相当于人类青春期前,n=6),12周龄的ICR雄性小鼠作为性成熟期组(相当于人类青壮年,n=6),12月龄的ICR雄性小鼠作为老年组(相当于人类中老年,n=6)。Real time PCR和Western Blot分别检测小鼠睾丸中Nur77 mRNA和蛋白水平表达。体外培养小鼠睾丸间质细胞系MLTC-1,Nur77干涉腺病毒(Ad-siNur77)及空载腺病毒(Ad-CMV)转染MITC-1。放射免疫法检测MLTC-1细胞培养液上清的睾酮含量,Real time pCR和Western Blot检测StAR及HSD3B的表达。结果 Nur77的mRNA水平在青春期前表达量最高,蛋白水平则在性成熟期小鼠睾丸中最高;Nur77干涉后MLTC-1细胞的睾酮合成含量较末干涉组明显降低;Nur77干涉后MLTC-1细胞的StAR和HSD3B的表达较未干涉组明显降低。结论 Nur77在小鼠睾丸组织的表达随年龄变化而变化,这种变化对于维持小鼠体内的睾酮水平可能有重要意义。
Objective To observe the expression of nuclear receptor Nur77 in testis of mice at different developmental stages and to explore its effect on testosterone synthesis. Methods ICR male mice of 1 week old were used as premature development group (equivalent to pre-puberty, n = 6) in the juvenile group (equivalent to human juvenile age, n = 6) 12-week-old ICR male mice served as the sex maturation group (equivalent to human young adults, n = 6), and 12-month-old ICR male mice as the elderly group (equivalent to middle-aged and elderly humans, n = 6). Real time PCR and Western Blot were used to detect the expression of Nur77 mRNA and protein in mouse testis respectively. The mouse testicular stromal cell line MLTC-1, Nur77 interference adenovirus (Ad-siNur77) and Ad-CMV were transfected into MITC-1 cells in vitro. The levels of testosterone in MLTC-1 cell culture supernatant were detected by radioimmunoassay. The expression of StAR and HSD3B were detected by Real time pCR and Western Blot. Results The mRNA level of Nur77 was highest before puberty and the protein level was the highest in testes of sexual maturity mice. The level of testosterone synthesis in MLTC-1 cells was significantly decreased after Nur77 intervention. The expression of StAR and HSD3B were significantly lower than those in the non-intervention group. Conclusion The expression of Nur77 in mouse testis varies with age, and this change may be of great significance for the maintenance of testosterone in mice.